Abstract Jaboticaba peels are an important source of health-benefit and antimicrobial compounds. The present work aims to evaluate concentration of polyphenolics and the mode of action of aqueous extract from jaboticaba peels against Staphylococcus aureus and Escherichia coli. Jaboticaba peel extract showed minimum inhibitory concentration and minimum bactericidal concentration against S. aureus of 5.1 g L-1 and 10.1 g L-1, respectively; meanwhile, against E. coli the parameters were 2.0 g L-1 and 3.4 g L-1. Kinetics of viable cell counts indicated a bacteriolytic action against both bacteria and Scanning Electron Microscopy (SEM) showed that jaboticaba peel extract causes subtle morphological changes in bacterial cells. Concentration of total polyphenols in the extract was 1535.04±36.05 mg of gallic acid equivalent (GAE) mL-1, monomeric anthocyanins was 14.52 ± 0.98 mg of cyanidin 3-glucoside mL-1, condensed tannins was 0.49 ± 0.05 mg of epicatechin equivalent mL-1 and phenolic acids was 80.04 ± 4.11 mg of caffeic acid equivalent (CAE) mL-1, which have demonstrated well-documented antibacterial activity. In conclusion, jaboticaba peel aqueous extract may be an interesting natural preservative to control Gram-positive and Gram-negative bacteria growth when interacting with the bacteria cell wall.

Resumo As cascas de jabuticaba são uma importante fonte de benefícios à saúde e compostos com atividade antimicrobiana. O presente trabalho tem como objetivo avaliar a concentração de polifenóis e o modo de ação do extrato aquoso da casca da jabuticaba contra Staphylococcus aureus e Escherichia coli. O extrato da casca da jabuticaba apresentou concentração inibitória mínima e concentração bactericida mínima contra S. aureus de 5,1 g L-1 e 10,1 g L-1, respectivamente; já contra E. coli, esses parâmetros foram 2,0 g L-1 e 3,4 g L-1. A cinética da contagem de células viáveis indica uma ação bacteriolítica contra ambas as bactérias e a microscopia eletrônica de varredura mostrou que o extrato da casca da jabuticaba causa sutis alterações morfológicas nas células bacterianas. A concentração de polifenóis totais no extrato foi 1.535,04±36,05 mg ácido gálico equivalente mL-1; antocianinas monoméricas, 14,52±0,98 mg cianidina 3-glicosídeo mL-1; taninos condensados, 0,49 ± 0,05 mg equivalente epicatequina mL-1, e ácidos fenólicos, 80,04±4,11 mg ácido cafeico equivalente mL-1, compostos que já demonstraram atividade antibacteriana bem documentada. Em conclusão, o extrato aquoso da casca de jabuticaba mostra-se um bioconservante interessante para o controle de bactérias Gram-positivas e Gram-negativas, e interage com a parede celular bacteriana.

Abstract Gut microbiota exerts a fundamental role in human health and increased evidence supports the beneficial role of probiotic microorganisms in the maintenance of intestinal health. Enterococcus durans LAB18S was previously isolated from soft cheese and showed some desirable in vitro probiotic properties, for that reason its genome was sequenced and evaluated for genes that can be relevant for probiotic activity and are involved in selenium metabolism. Genome sequencing was performed using the Illumina MiSeq System. A variety of genes potentially associated with probiotic properties, including adhesion capability, viability at low pH, bile salt resistance, antimicrobial activity, and utilization of prebiotic fructooligosaccharides (FOS) were identified. The strain showed tolerance to acid pH and bile salts, exhibited antimicrobial activity and thrived on prebiotic oligosaccharides. Six genes involved in selenium metabolism were predicted. Analysis of the SECIS element showed twelve known selenoprotein candidates. E. durans LAB18S was the only food isolate showing absence of plasmids, virulence and antimicrobial resistance genes, when compared with other 30 E. durans genomes. The results of this study provide evidence supporting the potential of E. durans LAB18S as alternative for probiotic formulations.

Abstract The aim of this study was to investigate the combined effect of carvacrol, thymol and nisin against Staphylococcus aureus and the combined effect of carvacrol and thymol against Salmonella Enteritidis. Minimum inhibitory concentrations (MIC) of carvacrol, thymol, and nisin for S. aureus were 200, 150 and 30 µg/mL, respectively. MIC of carvacrol and thymol for Salmonella Enteritidis was 200 µg/mL. A factorial method of independent variables was then used to study the combined effect of antimicrobials. Results showed that combinations of carvacrol-thymol-nisin (reduction of 1.2 log CFU/mL for MIC and 4.98 log CFU/mL for 2MIC), carvacrol-thymol (reduction of 1.33 log CFU/mL for 2MIC), nisin-thymol (reduction of 3.52 log CFU/mL for 2MIC) and nisin-carvacrol (reduction of 3.41 log CFU/mL for 2MIC) attained a significant inhibition of S. aureus. Similarly, there was significant reduction of Salmonella Enteritidis due to combined effect of thymol-carvacrol (reduction of 4.5 log CFU/mL for MIC and inhibition below detection limit for 2MIC). Therefore, the combinations of natural antimicrobials described in this work showed potential to be used as an additional barrier for food safety.

Abstract Bacteria of the genus Bacillus sp. present the potential for inhibiting various pathogens, making them a promising starting point in the search for new antimicrobial substances. In this study, bacteria were isolated from sediment samples from humid areas of a Natural Conservation Unit in the state of Rio Grande do Sul, Brazil. The isolate Bacillus sp. sed 1.4 was selected for production of antimicrobial activity, and was characterized by MALDI-TOF and 16S rDNA sequencing. Phylogenetic analysis showed that Bacillus sed 1.4 was closely related to Bacillus altitudinis and Bacillus pumilus. The cell-free supernatant was partially purified using ammonium sulfate precipitation, gel filtration chromatography (Sephadex G-200) and an ultrafiltration membrane. Partial purification resulted in specific activity of 769.23 AU/mg, with a molecular mass of approximately 148 kDa. This antimicrobial substance showed stability at 100°C for 5 min, and was inactivated by proteolytic enzymes. An antimicrobial effect against Listeria species was observed. Considering the importance of the Listeria genus in the area of food safety, this antimicrobial activity should be further explored, specifically in the field of dairy products and with a focus on food biopreservation studies.

Resumo O objetivo deste trabalho foi desenvolver formulações de misturas para preparo de bolos de caneca, sem ovos e sem leite, com farinhas de cascas e sementes de uvas, além de avaliar características físico-químicas, tecnológicas e sensoriais, com enfoque nos efeitos dos ácidos ascórbico e cítrico sobre os bolos, com substituição parcial da farinha de trigo por farinha de bagaço de uva. Aos produtos, foram incorporados 5% de farinhas de cascas e sementes em relação à farinha de trigo. Foram realizadas análises de polifenóis, atividade antioxidante, cor, pH, volume específico, perda de massa e sensorial. Entre as formulações com farinha de uva, a amostra com adição de ácido ascórbico apresentou maior (p < 0,05) atividade antioxidante, medida pelo método ABTS, do que as amostras sem adição de ácidos. Entretanto, a formulação não apresentou diferença significativa (p > 0,05) quando comparada com a formulação com a adição de ácido cítrico ou a combinação de ácidos. De acordo com o coeficiente de correlação de Pearson, quanto mais ácido (menor pH) se mostrou o bolo, maior foi a tendência de aumento na concentração de polifenóis totais, antocianinas e compostos com atividade antioxidante. Com relação ao volume específico, a maioria das formulações controles apresentou os maiores volumes em relação às suas respectivas amostras com farinha de uva. Assim, a incorporação dos ácidos ascórbico e cítrico nos bolos com farinha de bagaço de uva possivelmente acarretou estabilização da coloração vermelha e o bolo com farinha de bagaço de uva e ácido ascórbico apresentou as maiores quantidades de compostos fenólicos totais, quando comparado aos outros bolos com o subproduto, além de ter sido um dos bolos mais bem aceitos pelos provadores, demonstrando o potencial de mercado para esses produtos alimentícios inovadores.

Abstract This study aimed to develop formulations of mixtures to prepare mug cakes, without eggs and milk, with grape skin and seed flours, and to evaluate their physicochemical, technological and sensory characteristics focusing on the effects of ascorbic and citric acids on cakes with partial replacement of wheat flour with grape marc flour. The products were incorporated with 5% grape marc flour in relation to wheat flour. The following analyses were performed: polyphenols, antioxidant activity, color, pH, specific volume, loss of mass, and sensory. Among the formulations with grape marc flour, the sample with addition of ascorbic acid showed higher (p < 0.05) antioxidant activity measured by the ABTS method than the samples without addition of acids. However, this formulation showed no statistically significant difference (p > 0.05) compared with those with addition of citric acid or a combination of acids. According to the Pearson’s correlation coefficient, the more acidic (lower pH) the cake was, the greater the tendency for increased concentration of total polyphenols, anthocyanins, and compounds with antioxidant activity. Regarding specific volume, most of the control formulations presented higher values compared with those of their respective samples with grape marc flour. Thus, incorporation of ascorbic and citric acids into cakes with grape marc flour possibly resulted in red color stabilization, and the cake with grape marc flour and ascorbic acid presented the highest amounts of total phenolic compounds compared with those of other cakes with this by-product, in addition to being one of the cakes most accepted by the tasters, demonstrating the market potential of these innovative food products.

Abstract Wetlands are ecosystems rich in biodiversity and their ecological importance is recognized worldwide. Sediment samples were subjected to physical-chemical analysis and organic carbon content varied from 3.0% to 4.8%, the clay between 32 and 40%, silt with 41% and 43%, sand coarse varied between 6 and 11% and fine sand between 7 and 16%. The nitrogen values varied from 0.25% to 0.48%, the pH from 5.4 to 7.5 and the humidity varied from 44 to 56%. The selected isolates were evaluated for enzymatic properties and 64% showed positive results for amylase, 16% for gelatinase, 37% for lipase, 91% for protease and 2.7% for inulinase. Six bacterial isolates were selected for the overlapping assay and Bacillus sp. sed 2.2 showed inhibitory activity against Corynebacterium fimi NCTC 7547, and the antimicrobial substance was partially purified. The characterization of the substance was carried and the substance was stable at 100° C for up to 10 minutes and sensitive to the enzymes papain and trypsin. This substance was active against some species of Listeria, including Listeria monocytogenes ATCC 7644. The microorganims obtained from sediment samples were important sources of bioactive compounds, including enzymes and peptides, being a source of bioactive compounds to be studied.

Nanostructured zinc oxide (ZnO) prepared by combustion in solution was used to obtain nanocomposites. The ZnO particles were characterized by Brunauer-Emmett-Teller (BET), X-ray diffraction (XRD), and scanning electron microscopy (SEM), showing crystallite size of 32 nm and a superficial area of 32.6 m2 g-1. Nanocomposites with 1, 3, and 5 wt.% of ZnO in the polymeric matrix were obtained using the in situ polymerization of ethylene with catalytic activities between 1500-1700 kg (molZr h PE)-1. The high-density polyethylene nanocomposites (PEZnO) were characterized by thermogravimetric analysis (TGA), differential scanning calorimetry (DSC), SEM, and transmission electron microscopy (TEM). The nanocomposites with 1 wt.% ZnO gave excellent mechanical properties, and all were active against Gram-positive (Staphylococcus aureus) and Gram-negative (Escherichia coli) bacteria.

ABSTRACT The adhesion ability of bacteria to abiotic surfaces has important implications in food industries, because these organisms can survive for long periods through the biofilm formation. They can be transferred from one place to another in the industry causing contamination of the food processing environment. In this study, the antibacterial and antibiofilm activities of the antimicrobial peptide P34, characterized as a bacteriocin-like substance (BLS P34) were tested against planktonic and sessile cells of Staphylococcus aureus and Enterococcus faecalis isolated from foods. The BLS P34 showed inhibitory effect against all planktonic cells of E. faecalis. The inhibition of biofilm formation and the eradication of pre-formed biofilm were evaluated with the crystal violet assay and with the reduction of 3-bromide [4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium. The BLS P34 promoted a reduction of percentage of adhered microbial cells on the surface, not being able to perform the complete elimination of biofilm formation. The metabolic activity of S. aureus biofilms decreased considerably between 41-95%. However, E. faecalis cells showed up metabolically stimulated. The BLS P34 has the potential antibiofilm for the species S. aureus. Studies suggest more detailed approaches to a better understanding of the interactions between the antimicrobial and bacterial cells within the biofilm structure.

RESUMO: A atividade antimicrobiana do peptídeo P34 já foi previamente demonstrada. Neste estudo, o potencial antiviral do P34 e o mecanismo de ação in vitro contra o alfaherpesvírus bovino tipo 1 (BoHV1) foram investigados. O P34 exibiu baixa toxicidade, alto índice de seletividade (22.9) e percentagem de inibição viral de até 100% em células MDBK. Os resultados dos ensaios antivirais indicaram que não interage com receptores celulares, mas é capaz de inibir a penetração viral, imediatamente após a pré-adsorção. Além disso, a curva de crescimento do BoHV1 em células MDBK na presença do P34 revelou uma significativa redução no título somente após 8h de infecção, sugerindo também uma importante atividade do peptídeo nas fases finais do ciclo replicativo. Efeito virucida frente / BoHV1 foi observado apenas em concentrações citotóxicas do peptídeo. Os dados obtidos indicam que o peptídeo antimicrobiano P34 pode ser considerado um potencial composto inibidor de herpesvírus, in vitro, e estimulam posteriores investigações sobre sua atividade anti-herpética em modelos animais, bem como contra outros vírus.

ABSTRACT: Previous studies have demonstrated the antimicrobial activity of the peptide P34. In this study, the antiviral potential of P34 and the in vitro mechanism of action were investigated against bovine alphaherpesvirus type 1 (BoHV1). P34 exhibited low toxicity, a high selectivity index (22.9) and a percentage of inhibition of up to 100% in MDBK cells. Results from antiviral assays indicated that P34 did not interact with cell receptors, but it was able to inhibit the viral penetration immediately after pre-adsorption. In addition, BoHV1 growth curve in MDBK cells in the presence of P34 revealed a significant reduction in virus titer only 8h post-infection, also suggesting an important role at late stages of the replicative cycle. Virucidal effect was observed only in cytotoxic concentrations of the peptide. These findings showed that the antimicrobial peptide P34 may be considered as a potential novel inhibitor of in vitro herpesviruses and must encourage further investigation of its antiherpetic activity in animal models as well as against a wide spectrum of viruses.

RESUMO O yacon é uma raiz que contém compostos com atividade antioxidante e prebiótica. Este estudo teve como objetivo avaliar as características físicas, químicas e prebióticas das micropartículas de yacon encapsuladas por atomização. A partir do extrato do yacon foram obtidas as soluções de retido e permeado usando a separação por ultrafiltração (UF) com membranas de 10 e 30 kDa. O permeado foi misturado com goma arábica em concentrações de 10 e 15% e encapsulado por atomização a temperaturas de 140 e 160 °C. As micropartículas obtidas tiveram valores de solubilidade maiores que 90%, enquanto que a higroscopicidade diminuiu com o aumento da concentração de goma e da temperatura de secagem. A microscopia eletrônica mostrou uma forte tendência à aglomeração de partículas menores em torno das maiores, principalmente a 140 ºC. Em relação à cor, o parâmetro L* mostrou que a secagem a 160 ºC produziu amostras mais escuras, e os parâmetros a* e b* indicaram que todas as amostras tiveram coloração amarelo esverdeado. A concentração de inulina diminuiu durante a secagem, enquanto que os conteúdos de glicose e de frutose aumentaram devido às reações de termólise, o que levou à degradação das cadeias de inulina. As dispersões permeadas e retidas provenientes da separação de UF apresentaram atividade prebiótica, enquanto que somente o pó encapsulado proveniente da membrana de 30 kDa foi metabolizado pelo Lactobacillus acidophilus, apresentando escores de atividade sem diferença significativa em relação à da glicose.

ABSTRACT Yacon root is a functional food which contains antioxidants and prebiotics compounds. This study aimed to evaluate the physical, chemical and prebiotic characteristics of a yacon extract powder obtained by ultrafiltration (UF) with membranes of 10 and 30 kDa and encapsulation of the resulting permeate by spray drying. Drying air temperatures of 140 and 160 ºC and concentrations of gum arabic of 10 and 15% were tested. The samples had solubility values greater than 90% while the hygroscopicity decreased with increasing gum concentration and drying temperature. Electron microscopy showed a strong tendency to agglomeration of smaller particles around the larger ones, mainly at a temperature of 140 ºC. Regarding color, the parameter L* showed that drying at 160 ºC produced darker samples and the parameters a* and b* indicated that all samples were greenish yellow. The concentration of inulin decreased during drying, whereas the levels of glucose and fructose increased due to the thermolysis reaction, which led to degradation of inulin chains at drying temperature. The permeates and retentates from the UF membranes had prebiotic activity, while only the encapsulated product from UF-30 membrane, metabolized by Lactobacillus acidophilus LA-5(r), presented activity scores without significant difference to that of glucose.

Abstract Pecan nutshell is a residue from food industry that has potential to be used as biopreservative in foods. The objective of this study was to evaluate the antimicrobial activity of pecan nutshell aqueous extract in vitro and its effectiveness to inhibit spoilage microorganisms on lettuce leaves. The results indicate that the aqueous extract presents inhibitory activity against important foodborne pathogenic bacteria such as Listeria monocytogenes, Salmonella Enteritidis, Staphylococcus aureus, Bacillus cereus, Aeromonas hydrophila and Pseudomonas aeruginosa. Antimicrobial activity was not observed against Corynebacterium fimi, Clostridium perfringens, Escherichia coli, and the phytopathogenic fungi tested. When applied onto lettuce leaves, pecan nutshell extract reduced the counts of mesophilic and psychrotrophic bacteria in 2 and 4 log CFU/g, respectively, during storage of leafy for 5 days at refrigeration temperature (5 °C). The extract was not effective to inhibit yeast on lettuce leaves. Thus, the aqueous extract of pecan shell showed great potential to be used as a natural preservative in foods, acting mainly in the inhibition of spoilage and pathogenic bacteria.

RESUMOAs raízes de yacon contém inulina, o qual possui propriedade prebiótica e pode ser usado como substituto de sacarose ou gordura. Entretanto, a inulina é solúvel em água. As perdas desse nutriente durante o branqueamento são causadas, principalmente, por difusão ou lixiviação, o qual pode ser diminuído se as condições de tempo e temperatura são corretamente empregadas. Objetivou-se neste trabalho, foi estudar a lixiviação, durante o branqueamento em água quente em diferentes condições de tempo e temperatura, dos açúcares inulina, glicose e frutose presentes na raiz do yacon. As amostras foram previamente limpas, descascadas e cortadas na forma geométrica de disco de 1,75 ± 0,35 milímetros de espessura. Os resultados indicaram que o tempo e a temperatura foram significativos na dissolução dos açúcares. As menores perdas de inulina ocorreram à temperaturas e tempos menores que 60 ºC e 3 minutos, respectivamente, enquanto que as menores perdas de glicose e frutose, para todas as temperaturas, foram obtidas para tempos menores que 3 e 5 minutos, respectivamente.

ABSTRACTYacon roots contain inulin, which has prebiotic properties and it may be used as sucrose or fat substitutes. However, inulin is very soluble in water. The loss of this important nutrient during blanching is caused mainly by diffusion or leaching, which might be diminished if blanching temperature - time conditions are correctly employed. The aim of this study was to determine the leaching of the sugars inulin, glucose and fructose, present in yacon roots, during hot water blanching under different time/temperature conditions. The samples were cleaned and peeled and cut into geometric forms of 1.75 ± 0.35 mm thick disks. A complete factorial experimental design was used, and the treatments of the samples were compared using the Tukey test. The results indicated that the time and temperature were significant in the dissolution of the sugars. The lowest inulin losses occurred at temperatures and times lower than 60 ºC and 3 minutes. For all temperatures, the lowest glucose and fructose losses were obtained at time lower than 3 and 5 minutes, respectively.

P34 is an antimicrobial peptide produced by a Bacillus sp. strain isolated from the intestinal contents of a fish in the Brazilian Amazon basin with reported antibacterial activity. The aim of this work was to evaluate the peptide P34 for its in vitro antiviral properties against canine adenovirus type 2 (CAV-2), canine coronavirus (CCoV), canine distemper virus (CDV), canine parvovirus type 2 (CPV-2), equine arteritis virus (EAV), equine influenza virus (EIV), feline calicivirus (FCV) and feline herpesvirus type 1 (FHV-1). The results showed that the peptide P34 exhibited antiviral activity against EAV and FHV-1. The peptide P34 inhibited the replication of EAV by 99.9% and FHV-1 by 94.4%. Virucidal activity was detected only against EAV. When P34 and EAV were incubated for 6 h at 37 °C the viral titer reduced from 10(4.5) TCID50 to 10(2.75) TCID50, showing a percent of inhibition of 98.6%. In conclusion, our results demonstrated that P34 inhibited EAV and FHV-1 replication in infected cell cultures and it showed virucidal activity against EAV. Since there is documented resistance to the current drugs used against herpesviruses and there is no treatment for equine viral arteritis, it is advisable to search for new antiviral compounds to overcome these infections.

The antimicrobial activity of the bacteriocin-like substance (BLS) P34 against Listeria monocytogenes was investigated in chicken sausage. The BLS was applied to chicken sausages (256 AU g-1) previously inoculated with a suspension of 10² cfu g-1 of L. monocytogenes. BLS P34 inhibited the indicator microorganism in situ in all incubation times for up to 10 days at 5 °C. The effectiveness of BLS P34 was increased when it was added in combination with nisin. The bacteriocin was also tested in natural eatable natural bovine wrapping (salty semi-dried tripe) against the same indicator microorganism, also showing inhibitory capability in vitro. BLS P34 showed potential to control L. monocytogenes in refrigerated meat products.

Prolonged culturing of many microorganisms leads to the loss of virulence and a reduction of their infective capacity. However, little is known about the changes in the pathogenic strains of Acanthamoeba after long culture periods. Our study evaluated the effect of prolonged culturing on the invasiveness of different isolates of Acanthamoeba in an in vivo rat model. ATCC strains of Acanthamoeba, isolates from the environment and clinical cases were evaluated. The in vivo model was effective in establishing the infection and differentiating the pathogenicity of the isolates and re-isolates. The amoebae cultured in the laboratory for long periods were less virulent than those that were recently isolated, confirming the importance of passing Acanthamoeba strains in animal models.